Cellometer for Analyzing NCI-60 Cancer Cell Lines

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Introduction to NCI-60 Human Cancer Cell Lines

The late 1980s saw the development and creation of the National Cancer Institute (NCI) 60 human tumor
cell line anticancer drug discovery project. This project was intended to screen thousands of pharmacological
compounds for anticancer activity, thus replacing earlier cumbersome en vivo models. The 60 created cell lines
represent nine human cancers: breast, central nervous system, colon, kidney, leukemia, lung, melanoma, ovary,
and prostate. Since its inception, the testing of anticancer drugs using the NCI-60 panel has generated the
most extensive cancer pharmacology database in the world.3 It has also provided invaluable knowledge in genome mutations, drug –
tumor interactions and general tumor biology.

Here we have performed Cellometer image cytometry on all NCI-60 cancer cell lines. Every cell sample was
individually stained with trypan blue, imaged and analyzed using the Cellometer T4. Each sample was assessed for
viability and cell size. The ability to accurately measure cell viability and cell size are two important components
in determining the functionality anticancer compounds. Cell death, or reduced cell size in certain tumor cell
lines could be an indicator of compound efficacy.

Measuring Cell Concentration and Viability

Simple, User-friendly Cellometer Procedure

After staining the cells with trypan blue, and loading the sample into the counting chamber slide, the slide is inserted into the Cellometer
instrument which automatically captures and analyzes multiple cell images. Within seconds, the result table
displays the live, dead, and total cell concentrations, as well as the percent viability.

Measuring Cell Size of NCI-60 Cancer Cell Lines

The ability to measure cell size has been a valuable tool for researchers examining the effects of
pharmacological agents on cancer cells. Many of the NCI-60 cancer cell lines have distinctly different
morphological characteristics and cell sizes.

The Cellometer uses an "Accurate Cell Membrane Outline Algorithm" that produces the cell membrane outline
around each cell. When the counting is complete, a green outline can be seen around each counted cell.
This sophisticated algorithm allows for consistent identification of the desired cell population and
provides reliable and accurate cell counts, concentration, viability and cell size measurements. After
each count a Cell Diameter Histogram is automatically generated representing the cell size distribution
of the counted population.

786-O cells

Trypan Blue stained 786-O cells

Cellometer counted 786-O cells

In this experiment average cell diameter for 786-O cells was measured to be 17.6 microns. From the cell diameter histogram, we can estimate that approximately 60%
of the cells had a cell diameter that was between 16.6 and 18.6 microns.

SW-620 cells

Trypan Blue stained SW-620 cells

Cellometer counted SW-620 cells

In comparison to 786-O cells, the average diameter of SW-620 cells was measured to be 11.1 microns. Looking at the cell diameter histogram, approximately 75 % of the cells had a cell diameter that was between 10.5 and 12.0 microns.

NCI-60 Human Cancer Cell Lines

Using the Cellometer Auto T4, we imaged and counted every NCI-60 cancer cell line, obtaining the
cell concentration, viability, and cell size. Shown here, are the enlarged representative images of
each cell line. Since all of the cells were stained with trypan blue, we show the stained trypan blue
image as well as the Cellometer counted image.

The cells are organized alphabetically by the organ or origin.

Breast

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

BT-549 cells

Hs 578T

MCF7

MDA-MB-231

MDA-MB-468

T-47D

Central Nervous System

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

SF-268

SF-295

SF-539

SNB-19

SNB-75

U251

Colon

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

Colo205

HCC-2998

HCT-116

HCT-15

HT29

KM12

SW-620

Kidney

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

786-O

A498

ACHN

CAKI-1

RXF 393

SN12C

TK-10

UO-31

Leukemia

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

CCRF-CEM

HL-60

K-562

MOLT-4

RPMI-8226

SR

Lung

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

A549

EKVX

HOP-62

HOP-92

NCI-H226

NCI-H23

NCI-H322M

NCI-H460

NCI-H522

Melanoma

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

LOX-IMVI

M14

MALME-3M

MDA-MB-435

SK-MEL-2

SK-MEL-28

SK-MEL-5

UACC-257

UACC-62

Ovarian

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

IGROV1

NCI/ADR-RES

OVCAR-3

OVCAR-4

OVCAR-5

OVCAR-8

SK-OV-3

Prostate

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

Conclusions

The Cellometer line of instruments is capable of measuring the cell concentration, viability, and cell size of all 60 NCI cancer cell lines. The innovative "Accurate Cell Membrane Outline Algorithm" assures that all cell samples are de-clustered and counted properly. The ability of the Cellometer instruments to accurately and consistently measure cell concentration, viability, and cell size provides researchers with a reliable tool in their cancer research.

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